This invention relates to therapeutic uses, methods, compounds, formulations, drinks and food stuffs involving, containing, comprising and/or including certain isoflavone compounds.
The isoflavone compounds according to this invention are described by general formula I, 
in which
Z is H,
R1 is H, or RACO where RA is C1-10 alkyl or an amino acid,
R2 is H, OH, or ORB where RB is an amino acid or CORA where RA is as previously defined, W is H, A is H or OH, and B is selected from 
W is H, and A and B taken together form a six membered ring selected from 
W, A and B taken with the groups with which they are associated comprise 
W and A taken together with the groups with which they are associated comprise 
and B is 
wherein
R3 is H, CORA where RA is as previously defined, CO2RC where RC is C1-10 alkyl, or CORB where RB is as previously defined,
R4 is H, CORD where RD is H, OH, C1-10 alkyl or an amino acid, CO2RC where RC is as previously defined, CORE where RE is H, C1-10 alkyl or an amino acid, COOH, CORC where RC is as previously defined, or CONHRE where RE is as previously defined,
R5 is H, CO2RC where RC is as previously defined, or CORCORE where RC and RE are as previously defined, and where the two R5 groups are attached to the same group they are the same or different,
R6 is H or hydroxy C1-10 alkyl,
X is preferably O, but may be N or S, and
Y is 
where R7 is H, or C1-10 alkyl.
provided that compounds of the formula I where
R1, W and Z are H
R2 is H or OH
A and B taken together are a six membered ring 
Y is as defined above
and R7 is H or OCH3,
are specifically excluded, and
provided that where the method is a method for the treatment or prophylaxis of menopausal syndrome or premenstrual tension syndrome compounds of the formula I where
R1 is H, W and Z are H, and R2 is H or OH,
A and B taken together are a six membered ring 
or A is OH and B is 
where Y is as defined above and R7 is H are specifically excluded, and further provided that where the method is a method for the treatment of a form of cancer or rheumatoid arthritis compounds of the formula I where:
R1 and W are H, Z is H or OCH3 and R2 is H or OH, and
A and B taken together are a six membered ring of the formula 
Y is as defined above, R4 is CO2H or CO2CH2CH3, and R7 is H are specifically excluded.
Preferably the compounds of the formula I are selected from: 
wherein
R8 is COR where RD is as previously defined,
R9CO2RC or CORE where RC and RE are as previously defined,
R10 is CORC or CORCORE where RC and RE are as previously defined,
R11 is H or OH,
R12 is H, COOH, CO2RC where RC and is as previously defined, or CONHRE where RE is as previously defined,
R13 is OH, ORB where RB is as previously defined, or CORA where RA is as previously defined,
R14 is H, or CORA where RA is as previously defined,
R15 is CORA where RA is as previously defined,
R16 is H, CORB or CO2RC where RB and RC are as previously defined,
R17 is H or hydroxy C1-10 alkyl,
R18 is H or C1-10 alkyl,
and xe2x80x9c---xe2x80x9d represents either a single bond or a double bond.
Alkyl groups may be straight or branched chains. C1-10 alkyl preferably contains from 1 to 5 carbons, more preferably methyl, ethyl or propyl.
Certain of the above compounds may be referred to by the names dihydrodaidzein (compound 1 where R8 is H), dihydrogenestein (compounds 2 and 5), dehydro-O-desmethylangolensin (compound 11), tetrahydrodaidzein (compound 8), equol and dehydroequol (compound 10), O-desmethyl-angolensin (ODMAxe2x80x94compound 13), and 6-hydroxy-O-desmethylangolensin (6-hydroxy-ODMAxe2x80x94compound 14).
It has surprisingly been found by the inventors that compounds of the formula I, and more specifically compounds of the formulae 1 to 19 have particular utility and effectiveness in the treatment, prophylaxis, amelioration defence against, and/or prevention of menopausal syndrome including hot flushes, anxiety, and depression, mood swings, night sweats, headaches, and urinary incontinence; osteoporosis; premenstrual syndrome, including fluid retention, cyclical mastalgia, and dysmenorrhoea; Reynaud""s Syndrome; Reynaud""s Phenomenon; Buergers Disease; coronary artery spasm; migraine headaches; hypertension; benign prostatic hypertrophy; breast cancer; uterine cancer; ovarian cancer; testicular cancer; large bowel cancer; endometrial cancer; prostatic cancer uterine cancer; atherosclerosis; Alzheimers disease; inflammatory diseases including inflammatory bowel disease, ulcerative colitis, Crohns disease; rheumatic diseases including rheumatoid arthritis; acne; baldness including male pattern baldness (alopecia hereditaria); psoriasis and diseases associated with oxidant stress including cancer, myocardial infarction stroke, arthritis, sunlight induced skin damage or cataracts.
According to a first aspect of the present invention there is provided a method for the treatment, prophylaxis, amelioration, defence against, and/or prevention of menopausal syndrome including hot flushes, anxiety, and depression, mood swings, night sweats, headaches, and urinary incontinence; osteoporosis; premenstrual syndrome, including fluid retention, cyclical mastalgia, and dysmenorrhoea; Reynaud""s Syndrome; Reynaud""s Phenomenon; Buergers Disease; coronary artery spasm; migraine headaches; hypertension; benign prostatic hypertrophy; breast cancer; uterine cancer; ovarian cancer; testicular cancer; large bowel cancer; endometrial cancer; prostatic cancer; uterine cancer; atherosclerosis; Alzheimers disease; inflammatory diseases including inflammatory bowel disease, ulcerative colitis, Crohns disease; rheumatic diseases including rheumatoid arthritis; acne; baldness including male pattern baldness (alopecia hereditaria); psoriasis and diseases associated with oxidant stress including cancer, myocardial infarction stroke, arthritis, sunlight induced skin damage or cataracts which comprises administering to a subject a therapeutically effective amount of one or more compounds of the formula I: 
where R1, R2, Z, W, A and B are as previously defined, either alone or in association with one or more pharmaceutically acceptable carriers and/or excipients.
Preferably, one or more compounds of the formulae 1 to 19 may be used in the treatment, prophylaxis, amelioration of menopausal syndrome including hot flushes, anxiety, and depression, mood swings, night sweats, headaches, and urinary incontinence; osteoporosis; premenstnial syndrome, including fluid retention, cyclical mastalgia, and dysmenorrhoea; Reynaud""s Syndrome; Reynaud""s Phenomenon; Buergers Disease; coronary artery spasm; migraine headaches; hypertension; benign prostatic hypertrophy; breast cancer; uterine cancer; ovarian cancer; testicular cancer; large bowel cancer; endometrial cancer; prostatic cancer; uterine cancer; atherosclerosis; Alzheimers disease; inflammatory diseases including inflammatory bowel disease, ulcerative colitis, Crohns disease; rheumatic diseases including rheumatoid arthritis; acne; baldness including male pattern baldness (alopecia hereditaria); psoriasis and diseases associated with oxidant stress including cancer, myocardial infarction stroke, arthritis, sunlight induced skin damage or cataracts (for convenience hereafter referred to as the xe2x80x9ctherapeutic indicationsxe2x80x9d). Cancer, myocardial infarction, stroke, arthritis, sunlight induced skin damage and cataracts are generally regarded to be associated with oxidant stress. This invention includes the treatment of diseases associated with oxidant stress.
A second aspect of the present invention is the use of compounds of the formula I for the manufacture of a medicament for the treatment, amelioration, defence against, prophylaxis and/or prevention of one or more of the therapeutic indications. It is particularly preferred that one or more compounds of the formulae 1 to 19 are employed in the treatment, prophylaxis, amelioration, defence against, and/or prevention of said indications.
A third aspect of the present invention is use of one or more compounds of the formula I in the treatment, amelioration, defence against, prophylaxis and/or prevention of one or more of the therapeutic indications. Compounds of the formulae 1 to 19 are particularly preferred.
A fourth aspect of the present invention comprises an agent for the treatment, prophylaxis, amelioration, defence against and/or treatment of the therapeutic indications which comprises one or more compounds of the formula I either alone or in association with one or more carriers or excipients. Compounds of the formulae 1 to 19 are particularly preferred.
A fifth aspect of the invention is a therapeutic composition which comprises one or more compounds of the formula I in association with one or more pharmaceutical carriers and/or excipients. It is preferred that the compositions comprise one or more compounds of the formulae 1 to 19.
A sixth aspect of the present invention is a drink or food-stuff, which contains one or more compounds of the formula I. Preferably the food stuff contains one or more compounds of the formulae 1 to 19.
A seventh aspect of the present invention is a microbial culture or a food-stuff containing one or more microbial strains which microorganisms produce one or more compounds of the formula I. Preferably said microorganisms produce one or more of the compounds of the formulae 1 to 19.
An eighth aspect of the present invention relates to one or more microorganisms which produce one or more compounds of the formula I. Preferably the microorganism is a purified culture, which may be admixed and/or administered with one or more other cultures which produce compounds of the formula I. The compounds of the formula I preferably are selected from one or more of compounds of the formulae 1 to 19.
In a further aspect this invention is directed to compounds of the formula I. Preferably said compounds comprise compounds of the formulae 1 to 19.
Compounds of the present invention have particular application in the treatment of diseases associated with or resulting from estrogenic effects androgenic effects, vasolidatory and spasmodic effects, inflammatory effects and oxidative effects.
The amount of the compound of the formula I which is required in a therapeutic treatment according to the invention will depend upon a number of factors, which include the specific application, the nature of the particular compound used, the condition being treated, the mode of administration and the condition of the patient. In general, a daily dose per patient is in the range of 0.1 mg to 2 g; typically from 0.5 mg to 1 g; preferably from 50 mg to 200 mg.
Compounds of the formula I may be in a manner and amount as is conventionally practised. See, for example, Goodman and Gilman, The Pharmacological Basis of Therapeutics, 1299 (7th Edition, 1985). The specific dosage utilised will depend upon the condition being treated, the state of the subject, the route of administration and other well known factors as indicated above.
The production of a pharmaceutical composition for the treatment of the therapeutic indications herein described (for convenience hereafter referred to as the xe2x80x9cactive compoundsxe2x80x9d) are typically admixed with one or more pharmaceutically or veterinarially acceptable carriers and/or excipients as are well known in the art.
The carrier must, of course, be acceptable in the sense of being compatible with any other ingredients in the formulation and must not be deleterious to the subject. The carrier or excipient may be a solid or a liquid, or both, and is preferably formulated with the compound as a unit-dose, for example, a tablet, which may contain from 0.5% to 59% by weight of the active compound, or up to 100% by weight of the active compound. One or more active compounds may be incorporated in the formulations of the invention, which may be prepared by any of the well known techniques of pharmacy consisting essentially of admixing the components, optionally including one or more accessory ingredients.
The formulations of the invention include those suitable for oral, rectal, optical, buccal (for example, sublingual), parenteral (for example, subcutaneous, intramuscular, intradermal, or intravenous) and transdermal administration, although the most suitable route in any given case will depend on the nature and severity of the condition being treated and on the nature of the particular active compound which is being used.
Formulation suitable for oral administration may be presented in discrete units, such as capsules, cachets, lozenges, or tablets, each containing a predetermined amount of the active compound; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil emulsion. Such formulations may be prepared by any suitable method of pharmacy which includes the step of bringing into association the active compound and a suitable carrier (which may contain one or more accessory ingredients as noted above). In general, the formulations of the invention are prepared by uniformly and intimately admixing the active compound with a liquid or finely divided solid carrier, or both, and then, if necessary, shaping the resulting mixture such as to form a unit dosage. For example, a tablet may be prepared by compressing or moulding a powder or granules containing the active compound, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing, in a suitable machine, the compound of the free-flowing, such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, and/or surface active/dispersing agent(s). Moulded tablets may be made by moulding, in a suitable machine, the powdered compound moistened with an inert liquid binder.
Formulations suitable for buccal (sublingual) administration include lozenges comprising the active compound in a flavoured base, usually sucrose and acacia or tragacanth; and pastilles comprising the compound in an inert base such as gelatin and glycerin or sucrose and acacia.
Compositions of the present invention suitable for parenteral administration conveniently comprise sterile aqueous preparations of the active compounds, which preparations are preferably isotonic with the blood of the intended recipient. These preparations are preferably administered intravenously, although administration may also be effected by means of subcutaneous, intramuscular, or intradermal injection. Such preparations may conveniently be prepared by admixing the compound with water or a glycine buffer and rendering the resulting solution sterile and isotonic with the blood. Injectable formulations according to the invention generally contain from 0.1% to 60% w/v of active compound and are administered at a rate of 0.1 ml/minute/kg.
Formulations suitable for rectal administration are preferably presented as unit dose suppositories. These may be prepared by admixing the active compound with one or more conventional solid carriers, for example, cocoa butter, and then shaping the resulting mixture.
Formulations or compositions suitable for topical administration to the skin preferably take the form of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil. Carriers which may be used include Vaseline, lanoline, polyethylene glycols, alcohols, and combination of two or more thereof. The active compound is generally present at a concentration of from 0.1% to 0.5% w/w, for example, from 0.5% to 2% w/w. Examples of such compositions include cosmetic skin creams.
Formulations suitable for transdermal administration may be presented as discrete patches adapted to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. Such patches suitably contain the active compound as an optionally buffered aqueous solution of, for example, 0.1 M to 0.2 M concentration with respect to the said active compound.
Formulations suitable for transdermal administration may also be delivered by iontophoresis (see, for example, Pharmaceutical Research 3 (6), 318 (1986)) and typically take the form of an optionally buffered aqueous solution of the active compound. Suitable formulations comprise citrate or bis/tris buffer (pH 6) or ethanol/water and contain from 0.1 M to 0.2 M active ingredient.
The active compounds may be provided in the form of food stuffs, such as being added to, admixed into, coated, combined or otherwise added to a food stuff. The term food stuff is used in its widest possible sense and includes liquid formulations such as drinks including dairy products and other foods, such as health bars, desserts, etc. Food formulations containing compounds of the invention can be readily prepared according to standard practices.
Compounds of the present invention have potent antioxidant activity and thus find wide application in pharmaceutical and veterinary uses, in cosmetics such as skin creams to prevent skin ageing, in sun screens, in foods, health drinks, shampoos, and the like.
It has surprisingly been found that compounds of the formula I interact synergistically with vitamin E to protect lipids, proteins and other biological molecules from oxidation. Accordingly, a further aspect of this invention provides a composition comprising one or more compounds of the formula I, vitamin E, and optionally a pharmaceutically, veterinarially or cosmetically acceptable carriers and/or excipients.
Therapeutic methods, uses and compositions may be for administration to humans or animals, such as companion and domestic animals (such as dogs and cats), birds (such as chickens, turkeys, ducks), livestock animals (such as cattle, sheep, pigs and goats) and the like.
Compounds of the formula I may be prepared as follows:
A. Hydrogenation of daidzein, geniestein or derivatives thereof using palladium on calcium carbonate, as follows 
xe2x80x83where Axe2x80x2 is H or R1 where R1 is as previously defined and R8 and R11 and X are as previously defined. Compounds 2, 3, 4, 5, 6 and 7 may be produced by this method. Compounds 5 to 7 are enol forms of compounds 2 to 4.
B. Reduction of daidzein and daidzein derivatives with sodium borohydride as follows: 
xe2x80x83where R9, and X are as previously defined. Compound 8 may be produced by this method.
C. Hydrogenation of daidzein and diadzein derivatives using palladium on charcoal as a catalyst. 
xe2x80x83where R11 and R12 are as previously defined. Compound 10 may be produced by this method.
D. Acylation or resorcinol or derivatives thereof, followed by dehydrogenation with lithium bromide 
xe2x80x83Compounds 11 and 14 may be produced by this method. Compound 12 may be produced in a similar manner.
E. Acylation of 1, 3 and 5 trisubstituted benze with 4-hydroxyphenyl isopropyl acid or derivatives thereof 
xe2x80x83where R13 and R15 are as previsouly defined. Compounds 15 and 16 may be produced by this method.
F. Compounds of the formulae 17, 18 and 19 may be prepared according to the following reaction schemes.
(i) 
where R11, R17 and R18 are as previously defined.
G. HPLC fractionation of human urine/urine fractions of HPLC/GLC fractionation of bacterial culture supernatant so as to give purified compounds of the formulae 1 to 19. Product identity is confirmed by mas spectrometry. Compounds of the formulae 1 to 19 may be purified according to Joannou et al (1995) J. Steroid. Biochem. Molec. Biol. 54, 167-184, which is incorporated herein by reference.
It has surprisingly been observed by the inventors that the presence of isoflavonoids, in bodily secretions, more particularly, isoflavonoid metabolites in the urine of subjects, is associated with a specific therapeutic response, medical condition, or absence of a specific medical condition. Determining the specific biological fingerprint of different isoflavonoids excreted by individuals enables therapeutic methods of treatment to be carried out.